Autotrophic Plants Research Articles

Localised photosynthetic pigments in the node of a holoparasitic plant: support for shoot growth?

Understanding the role of photosynthesis in the holoparasitic genus Cuscuta may give insights into the biology and evolution of these plants. We hypothesised that light irradiance on the node of C. racemosa is important for sustaining the growth of the new shoot through photosynthesis. We blocked light irradiance on the node of detached twigs of C. racemosa for one week and measured twig fresh and dry weight, shoot length and photosynthetic pigment content. Even though fresh and dry weight did not differ between non-irradiated and irradiated twigs, the shoots of non-irradiated nodes were significantly shorter. The non-irradiated plants showed higher amounts of chlorophyll a, b, and lycopene, and a lower concentration of β-carotene. Furthermore, the chlorophyll a/b ratio was lower than that of the control (irradiated) plants. Beyond corroborating our hypothesis, our results suggest that, when light is blocked on the node of C. racemosa, the holoparasite responds in a way similar to that of autotrophic plants under shaded conditions. Our findings suggest a new and previously unacknowledged role for the node of C. racemosa, which would be supporting shoot elongation through photosynthesis.

The History and Diversity of Rice Domestication as Resolved From 1464 Complete Plastid Genomes.

The plastid is an essential organelle in autotrophic plant cells, descending from free-living cyanobacteria and acquired by early eukaryotic cells through endosymbiosis roughly one billion years ago. It contained a streamlined genome (plastome) that is uniparentally inherited and non-recombinant, which makes it an ideal tool for resolving the origin and diversity of plant species and populations. In the present study, a large dataset was amassed by de novo assembling plastomes from 295 common wild rice (Oryza rufipogon Griff.) and 1135 Asian cultivated rice (Oryza sativa L.) accessions, supplemented with 34 plastomes from other Oryza species. From this dataset, the phylogenetic relationships and biogeographic history of O. rufipogon and O. sativa were reconstructed. Our results revealed two major maternal lineages across the two species, which further diverged into nine well supported genetic clusters. Among them, the Or-wj-I/II/III and Or-wi-I/II genetic clusters were shared with cultivated (percentage for each cluster ranging 54.9%∼99.3%) and wild rice accessions. Molecular dating, phylogeographic analyses and reconstruction of population historical dynamics indicated an earlier origin of the Or-wj-I/II genetic clusters from East Asian with at least two population expansions, and later origins of other genetic clusters from multiple regions with one or more population expansions. These results supported a single origin of japonica rice (mainly in Or-wj-I/II) and multiple origins of indica rice (in all five clusters) for the history of rice domestication. The massive plastomic data set presented here provides an important resource for understanding the history and evolution of rice domestication as well as a genomic resources for use in future breeding and conservation efforts.

The features of xylem tracheary elements in some herbaceous members of the family Convolvulaceae Horan.

Numerous narrow xylem tracheary elements (tracheids and vessels) are present in liana stems, along with a few wide vessels that perform the main water-conducting function. This trait, known as “vessel dimorphism”, has been identified in studies on water-conducting tissue in autotrophic plants, including a large number of perennial climbing plants and a number of annual vines. Information is lacking on the presence of vessel dimorphism in parasitic plants of the lianescent habit. In this study, we performed a structural analysis of stems in the autotrophic herbaceous vines of Convolvulus arvensis L. and Calystegia sepium (L.) R. Br., as well as in the parasitic vines of Cuscuta monogyna Vahl, Cuscuta planiflora Ten., Cuscuta approximata Bab., and Cuscuta campestris Yunck., of the family of Convolvulaceae Horan. The xylem of C. arvensis and C. sepium contains a few wide conductive elements and many narrow ones. This feature is typical of autotrophic climbing plants. Only narrow tracheary elements are present in the xylem of the parasitic vines of the genus of Cuscuta L. (dodders). The total number of the tracheary elements is an order of magnitude less in the dodders than it is in the autotrophic vines. It is possible that the autotrophic ancestor of dodders lost the characteristic feature of the xylem of climbing plants, known as vessel dimorphism, during its transition to the parasitic lifestyle.

Novel mycorrhizal cheating in a green orchid: Cremastra appendiculata depends on carbon from deadwood through fungal associations.

To date, there has been no robust evidence for the exploitation of saprotrophic non-rhizoctonia fungi by green plants, although some fully mycoheterotrophic orchids are known to exploit them, and mycoheterotrophic evolution has probably occurred through intermediate mixotrophic stages. We investigated the physiological ecology of a fully mycoheterotrophic species Cremastra aphylla and its photosynthetic sister species Cremastra appendiculata, which putatively exploit saprotrophic fungi. Their mycorrhizal partners and ultimate nutritional sources were determined using molecular, stable isotopic, and radiocarbon analysis. Both Cremastra aphylla and Cremastra appendiculata were consistently associated with wood-decaying Psathyrellaceae. In addition, both species were highly enriched in carbon-13 (13 C) and, to a less degree, in nitrogen-15 (15 N). The δ13 C and δ15 N values of Cremastra appendiculata were intermediate between those of Cremastra aphylla and those of autotrophic plants. All Cremastra appendiculata samples and two Cremastra aphylla samples exhibited elevated Δ14 C values due to the acquisition of carbon fixed in wood during the past decades (14 C-enriched bomb carbon). Our multifaceted evidence indicated that both species obtained carbon from deadwood via saprotrophic fungi. Our findings strongly suggest that mixotrophic relationships associated with wood-decaying fungi represent a novel evolutionary pathway for full mycoheterotrophy in orchids.

Root Endophytic Fungal Community and Carbon and Nitrogen Stable Isotope Patterns Differ among Bletilla Species (Orchidaceae).

Orchids of the genus Bletilla are well-known ornamental plants and sources of traditional medicine in Asia that rely on the symbiotic relationship with root endophytic fungi throughout their whole life cycle. However, little is known about their fungal partners, infection pattern, and pathways of carbon gain. We investigated carbon and nitrogen stable isotope patterns in different organs of three Bletilla species, identified the root endophytic fungal community composition, and determined mycorrhizal colonization rates. The three Bletilla species were comprised by a polyphyletic group which belongs to different trophic modes, such as saprotroph, pathotroph, and symbiotroph; however, the dominant species and their abundances varied among Bletilla spp. Mycorrhizal infection rates also varied among Bletilla species, with B. striata (65% ± 25%) being significantly higher than those of B. formosana (35% ± 16%) and B. ochracea (22% ± 13%). Compared with surrounding autotrophic plants, all Bletilla spp. were significantly enriched in 13C with B. striata to a significantly higher level than other two Bletilla species. Among different organs, stems had higher δ13C values, while leaves and flowers had higher δ15N and total N content values across all three species. Our results indicate that the symbiotic relationship of Bletilla and its root endophytic fungi is not strictly specific. Although mycorrhizal infection rates were highly variable, the three Bletilla species had the same infection pattern with hyphae penetrating the cortex cell by the pathway cell. Different Bletilla species have different strategies for C allocation among plant organs. These findings provide new insights into the ecological adaptation of orchids and will contribute to Bletilla germplasm conservation and sustainable utilization.

Plastid Gene Transcription: An Update on Promoters and RNA Polymerases.

Chloroplasts, the sites of photosynthesis and sources of reducing power, are at the core of the success story that sets apart autotrophic plants from most other living organisms. Along with their fellow organelles (e.g., amylo-, chromo-, etio-, and leucoplasts), they form a group of intracellular biosynthetic machines collectively known as plastids. These plant cell constituents have their own genome (plastome), their own (70S) ribosomes, and complete enzymatic equipment covering the full range from DNA replication via transcription and RNA processive modification to translation. Plastid RNA synthesis (gene transcription) involves the collaborative activity of two distinct types of RNA polymerases that differ in their phylogenetic origin as well as their architecture and mode of function. The existence of multiple plastid RNA polymerases is reflected by distinctive sets of regulatory DNA elements and protein factors. This complexity of the plastid transcription apparatus thus provides ample room for regulatory effects at many levels within and beyond transcription. Research in this field offers insight into the various ways in which plastid genes, both singly and groupwise, can be regulated according to the needs of the entire cell. Furthermore, it opens up strategies that allow to alter these processes in order to optimize the expression of desired gene products.

Evolutionary histories and mycorrhizal associations of mycoheterotrophic plants dependent on saprotrophic fungi

Mycoheterotrophic plants (MHPs) are leafless, achlorophyllous, and completely dependent on mycorrhizal fungi for their carbon supply. Mycorrhizal symbiosis is a mutualistic association with fungi that is undertaken by the majority of land plants, but mycoheterotrophy represents a breakdown of this mutualism in that plants parasitize fungi. Most MHPs are associated with fungi that are mycorrhizal with autotrophic plants, such as arbuscular mycorrhizal (AM) or ectomycorrhizal (ECM) fungi. Although these MHPs gain carbon via the common mycorrhizal network that links the surrounding autotrophic plants, some mycoheterotrophic lineages are associated with saprotrophic (SAP) fungi, which are free-living and decompose leaf litter and wood materials. Such MHPs are dependent on the forest carbon cycle, which involves the decomposition of wood debris and leaf litter, and have a unique biology and evolutionary history. MHPs associated with SAP fungi (SAP-MHPs) have to date been found only in the Orchidaceae and likely evolved independently at least nine times within that family. Phylogenetically divergent SAP Basidiomycota, mostly Agaricales but also Hymenochaetales, Polyporales, and others, are involved in mycoheterotrophy. The fungal specificity of SAP-MHPs varies from a highly specific association with a single fungal species to a broad range of interactions with multiple fungal orders. Establishment of symbiotic culture systems is indispensable for understanding the mechanisms underlying plant–fungus interactions and the conservation of MHPs. Symbiotic culture systems have been established for many SAP-MHP species as a pure culture of free-living SAP fungi is easier than that of biotrophic AM or ECM fungi. Culturable SAP-MHPs are useful research materials and will contribute to the advancement of plant science.

Temporal Variation in Community Composition of Root Associated Endophytic Fungi and Carbon and Nitrogen Stable Isotope Abundance in Two Bletilla Species (Orchidaceae).

Mycorrhizae are an important energy source for orchids that may replace or supplement photosynthesis. Most mature orchids rely on mycorrhizae throughout their life cycles. However, little is known about temporal variation in root endophytic fungal diversity and their trophic functions throughout whole growth periods of the orchids. In this study, the community composition of root endophytic fungi and trophic relationships between root endophytic fungi and orchids were investigated in Bletilla striata and B. ochracea at different phenological stages using stable isotope natural abundance analysis combined with molecular identification analysis. We identified 467 OTUs assigned to root-associated fungal endophytes, which belonged to 25 orders in 10 phyla. Most of these OTUs were assigned to saprotroph (143 OTUs), pathotroph-saprotroph (63 OTUs) and pathotroph-saprotroph-symbiotroph (18 OTUs) using FunGuild database. Among these OTUs, about 54 OTUs could be considered as putative species of orchid mycorrhizal fungi (OMF). For both Bletilla species, significant temporal variation was observed in the diversity of root endophytic fungi. The florescence and emergence periods had higher fungal community richness of total species and endemic species than did other periods. Both Bletilla species were dominated by Agaricomycetes and Basidiomycota fungi throughout the whole year; however, their abundances varied between two Bletilla species and among phenological stages. Meanwhile, the ranges of 13C and 15N natural abundance were also highly dynamic across all growth stages of Bletilla species. Compared with the surrounding autotrophic plants, significant 13C enrichments (ε13C) were found across all phenological stages, while significant 15N enrichment in the florescence period and strong 15N depletion during the fruiting period were found for both Bletilla species. We can deduce that both Bletilla species obtained carbon from root endophytic fungi during the whole year. Additionally, the temporal varying tendency of root endophytic fungal diversity was consistent with 13C enrichments, which was also accord with the nutritional requirement of plant.

Mechanisms of resistance and virulence in parasitic plant-host interactions.

Parasitic plants pose a major biotic threat to plant growth and development and lead to losses in crop productivity of billions of USD annually. By comparison with "normal" autotrophic plants, parasitic plants live a heterotrophic lifestyle and rely on water, solutes and to a greater (holoparasitic plants) or lesser extent (hemiparasitic plants) on sugars from other host plants. Most hosts are unable to detect an infestation by plant parasites or unable to fend off these parasitic invaders. However, a few hosts have evolved defense strategies to avoid infestation or protect themselves actively post-attack often leading to full or partial resistance. Here, we review the current state of our understanding of the defense strategies to plant parasitism used by host plants with emphasis on the active molecular resistance mechanisms. Furthermore, we outline the perspectives and the potential of future studies that will be indispensable to develop and breed resistant crops.

The USE OF INDICATORS OF PLANT PHOTOSYNTHETIC AND ENZYMATICE ACTIVITY IN BIOINDICATION STUDIES

In the context of increasing anthropogenic impact on the environment, the study of the mechanisms of plant adaptation is an actual problem. To diagnose ecological troubles, it is advisable to study the indicators characterizing metabolic processes in autotrophic plants. The change in the parameters of the photosynthetic apparatus and antioxidant system in silver birch (Betula pendula), European spruce (Picea abies) and dandelion (Taraxacum officinale L.), depending on the intensity of anthropogenic load, was studied. The pigment composition of plants and the activity of antioxidant enzymes can be used as markers of the anthropogenic pollution level of a territory.

Specialized mycorrhizal association between a partially mycoheterotrophic orchid Oreorchis indica and a Tomentella taxon.

The evolution of full mycoheterotrophy in orchids likely occurs through intermediate stages (i.e., partial mycoheterotrophy or mixotrophy), in which adult plants obtain nutrition through both autotrophy and mycoheterotrophy. However, because of its cryptic manifestation, partial mycoheterotrophy has only been confirmed in slightly more than 20 orchid species. Here, we hypothesized that Oreorchis indica is partially mycoheterotrophic, since (i) Oreorchis is closely related to leafless Corallorhiza, and (ii) it possesses clustered, multi-branched rhizomes that are often found in fully mycoheterotrophic orchids. Accordingly, we investigated the nutritional modes of O. indica in a Japanese subboreal forest by measuring the 13C and 15N abundances and by community profiling of its mycorrhizal fungi. We found that O. indica mycorrhizal samples (all 12 samples from four individuals) were predominantly colonized by a single OTU of the obligate ectomycorrhizal Tomentella (Thelephoraceae). In addition, the leaves of O. indica were highly enriched in both 13C and 15N compared with those of co-occurring autotrophic plants. It was estimated that O. indica obtained 44.4±6.2% of its carbon from fungal sources. These results strongly suggest that in the Oreorchis-Corallorhiza clade, full mycoheterotrophy evolved after the establishment of partial mycoheterotrophy, rather than through direct shifts from autotrophy.

The fate of chlorophyll in phytophagous insects goes beyond nutrition.

Chlorophyll (Chl) is a natural compound that is found in all autotrophic plants. Since phytophagous insects ingest the photosynthetically active material with the plant leaves, the question arises if and how herbivores deal with Chl and its degradation products. Here we review findings on Chl degradation in phytophagous insects and highlight the role of these ubiquitous plant metabolites for plant-feeding insects. Due to the anaerobic gut of many insects, the degradation is limited to the removal of the peripheral substituents, while the tetrapyrrole core remains intact. Proteins, such as red fluorescent protein, P252 (a novel 252-kDa protein), and chlorophyllide binding protein have been reported to occur in the insect gut and might be indirectly connected to Chl degradation. Besides of an nutritional value, e.g., by taking up Mg2+ ions or by sequestration of carbon from the phytol side chain, the Chl degradation products may serve the insect, after binding to certain proteins, as antimicrobial, antifungal, and antiviral factors. The protein complexes may also confer protection against reactive oxygen species. The antibiotic potential of proteins and degradation products does not only benefit phytophagous insects but also human being in medical application of cancer treatment for instance. This review highlights these aspects from a molecular, biochemical, and ecological point of view.

Cuscuta australis (dodder) parasite eavesdrops on the host plants’ FT signals to flower

Many plants use environmental cues, including seasonal changes of day length (photoperiod), to control their flowering time. Under inductive conditions, FLOWERING LOCUS T (FT) protein is synthesized in leaves, and FT protein is a mobile signal, which is able to travel to the shoot apex to induce flowering. Dodders (Cuscuta, Convolvulaceae) are root- and leafless plants that parasitize a large number of autotrophic plant species with varying flowering time. Remarkably, some dodder species, e.g., Cuscuta australis, are able to synchronize their flowering with the flowering of their hosts. Detailed sequence inspection and expression analysis indicated that the FT gene in dodder C. australis very likely does not function in activating flowering. Using soybean host plants cultivated under inductive and noninductive photoperiod conditions and soybean and tobacco host plants, in which FT was overexpressed and knocked out, respectively, we show that FT-induced flowering of the host is likely required for both host and parasite flowering. Biochemical analysis revealed that host-synthesized FT signals are able to move into dodder stems, where they physically interact with a dodder FD transcription factor to activate dodder flowering. This study demonstrates that FTs can function as an important interplant flowering signal in host-dodder interactions. The unique means of flowering regulation of dodder illustrates how regressive evolution, commonly found in parasites, may facilitate the physiological synchronization of parasite and host, here allowing the C. australis parasite to time reproduction exactly with that of their hosts, likely optimizing parasite fitness.

Reconstructing plastome evolution across the phylogenetic backbone of the parasitic plant genusCuscuta(Convolvulaceae)

AbstractParasitic plants have evolved to have reduced or completely lost ability to conduct photosynthesis and are usually characterized by sweeping morphological, physiological and genomic changes. The plastid genome (or plastome) is highly conserved in autotrophic plants and houses many key photosynthesis genes. This molecule is thus a useful system for documenting the genomic effects of a loss of autotrophy. Cuscuta (dodders) represents one of 12 independent transitions to a parasitic lifestyle in angiosperms. This near-cosmopolitan genus contains > 200 obligate parasitic species circumscribed in four subgenera: Grammica, Pachystigma, Cuscuta and Monogynella. With respect to photosynthesis, Cuscuta is a heterogeneous group, containing both hemi- and holoparasitic members that are, respectively, partially or entirely reliant on parasitism to meet their carbon budget. Plastomes in this genus have been reported to show a substantial degree of diversification in terms of length and gene composition. Considered together with well-understood phylogenetic relationships, this genus presents an opportunity for fine-scale comparisons among closely related species of heterotrophic plants. This research documents changes in sequence composition and structure that occurred as these plants evolved along the trophic spectrum by using multiple whole-plastome assemblies from each of the four subgenera. By ‘triangulating’ the positions of genomic changes, we construct a step-by-s’tep model of plastome evolution across the phylogenetic backbone of Cuscuta and highlight the remarkable retention of most photosynthetic genes in these parasitic plants.

Fourfold daily growth rate in multicellular marine alga Ulva meridionalis

Microalgae with high growth rates have been considered as promising organisms to replace fossil resources with contemporary primary production as a renewable source. However, their microscopic size makes it hard to be harvested for industrial applications. In this regard, multicellular macroalgae are more suitable for harvesting. Here, we show that Ulva meridionalis has the highest growth rate ever reported for a multicellular autotrophic plant. Contrasted to the known bloom-forming species U. prolifera growing at an approximately two-fold growth rate per day in optimum conditions, U. meridionalis grows at a daily rate of over fourfold. The high growth ability of this multicellular alga would provide the most effective method for CO2 fixation and biomass production.

Contrasting patterns of local adaptation along climatic gradients between a sympatric parasitic and autotrophic tree species.

Sympatric tree species are subject to similar climatic drivers, posing a question as to whether they display comparable adaptive responses. However, no study has explicitly examined local adaptation of co-occurring parasitic and autotrophic plant species to the abiotic environment. Here we test the hypotheses that a generalist parasitic tree would display a weaker signal of selection and that genomic variation would associate with fewer climatic variables (particularly precipitation) but have similar spatial patterns to a sympatric autotrophic tree species. To test these hypotheses, we collected samples from 17 sites across the range of two tree species, the hemiparasite Nuytsia floribunda (n=264) and sympatric autotroph Melaleuca rhaphiophylla (n=272). We obtained 5,531 high-quality genome-wide single nucleotide polymorphisms (SNPs) for M.rhaphiophylla and 6,727 SNPs for N.floribunda using DArTseq genome scan technology. Population differentiation and environmental association approaches were used to identify signals of selection. Generalized dissimilarly modelling was used to detect climatic and spatial patterns of local adaptation across climatic gradients. Overall, 322 SNPs were identified as putatively adaptive for the autotroph, while only 57 SNPs were identified for the parasitic species. We found genomic variation to associate with different sets of bioclimatic variables for each species, with precipitation relatively less important for the parasite. Spatial patterns of predicted adaptive variability were different and indicate that co-occurring species with disparate life history traits may not respond equally to selective pressures (i.e., temperature and precipitation). Together, these findings provide insight into local adaptation of sympatric parasitic and autotrophic tree species to abiotic environments.

Cheating in arbuscular mycorrhizal mutualism: a network and phylogenetic analysis of mycoheterotrophy

Although mutualistic interactions are widespread and essential in ecosystem functioning, the emergence of uncooperative cheaters threatens their stability, unless there are some physiological or ecological mechanisms limiting interactions with cheaters. In this framework, we investigated the patterns of specialization and phylogenetic distribution of mycoheterotrophic cheaters vs noncheating autotrophic plants and their respective fungi, in a global arbuscular mycorrhizal network with>25000 interactions. We show that mycoheterotrophy evolved repeatedly among vascular plants, suggesting low phylogenetic constraints for plants. However, mycoheterotrophic plants are significantly more specialized than autotrophic plants, and they tend to be associated with specialized and closely related fungi. These results raise new hypotheses about the mechanisms (e.g. sanctions, or habitat filtering) that actually limit the interaction of mycoheterotrophic plants and their associated fungi with the rest of the autotrophic plants. Beyond mycorrhizal symbiosis, this unprecedented comparison of mycoheterotrophic vs autotrophic plants provides a network and phylogenetic framework to assess the presence of constraints upon cheating emergences in mutualisms.

Isotopic evidence of arbuscular mycorrhizal cheating in a grassland gentian species.

All orchids and pyroloids are mycoheterotrophic at least in the early stage. Many species are predisposed to mycoheterotrophic nutrition even in the adult stage, due to the initial mycoheterotrophy during germination. Although other green plants, such as gentian species, also produce numerous minute seeds, whose germination may depend on fungal associations to meet C demands, physiological evidence for partial mycoheterotrophy in the adult stage is lacking for most candidate taxa. Here, we compared the natural abundances of 13C and 15N isotopes in the AM-associated gentian species Pterygocalyx volubilis growing in high-light-intensity habitats with those of co-occurring autotrophic C3 and C4 plants and AM fungal spores. We found that P. volubilis was significantly enriched in 13C compared with the surrounding C3 plants, which suggests the transfer of some C from the surrounding autotrophic plants through shared AM networks. In addition, the intermediate δ15N values of P. volubilis, between those of autotrophic plants and AM fungal spores, provide further evidence for partial mycoheterotrophy in P. volubilis. Although it is often considered that light deficiency selects partial mycoheterotrophy, we show that partial mycoheterotrophy in AM-forming plants can evolve even under light-saturated conditions. The fact that there have been relatively few descriptions of partial mycoheterotrophy in AM plants may not necessarily reflect the rarity of such associations. In conclusion, partial mycoheterotrophy in AM plants may be more common than hitherto believed.

Some mycoheterotrophic orchids depend on carbon from dead wood: novel evidence from a radiocarbon approach.

Mycoheterotrophic plants depend entirely on fungal associations for organic nutrients. While most mycoheterotrophic plants are associated with the mycorrhizal partners of surrounding green plants, some mycoheterotrophs are believed to obtain carbon from decaying litter or dead wood by parasitising saprotrophic fungi, based on culture experiments and 13 C and 15 N isotopic signatures. The carbon age (the time since carbon was fixed from atmospheric CO2 by photosynthesis) can be estimated by measuring the concentration of 14 C arising from the bomb tests of the 1950s and 1960s. Given that mycorrhizal fungi obtain photosynthate from their plant partners, and saprotrophic wood-decaying fungi obtain carbon from older sources, radiocarbon could represent a new and powerful tool to investigate carbon sources of mycoheterotrophic plants. We showed that the Δ14 C values of mycoheterotrophs exploiting ectomycorrhizal fungi were close to 0‰, similar to those of autotrophic plants. By contrast, the Δ14 C values of mycoheterotrophs exploiting saprotrophic fungi ranged from 110.7‰ to 324.8‰, due to the 14 C-enriched bomb carbon from dead wood via saprotrophic fungi. Our study provides evidence supporting that some mycoheterotrophic orchids depend on forest woody debris. Our study also indicates that radiocarbon could be used to predict the trophic strategies of mycoheterotroph-associated fungal symbionts.

Three-year pot culture of Epipactis helleborine reveals autotrophic survival, without mycorrhizal networks, in a mixotrophic species

Some mixotrophic plants from temperate forests use the mycorrhizal fungi colonizing their roots as a carbon source to supplement their photosynthesis. These fungi are also mycorrhizal on surrounding trees, from which they transfer carbon to mixotrophic plants. These plants are thus reputed difficult to transplant, even when their protection requires it. Here, we take profit of a successful ex situ pot cultivation over 1 to 3 years of the mixotrophic orchid Epipacis helleborine to investigate its mycorrhizal and nutrition status. Firstly, compared with surrounding autotrophic plants, it did not display the higher N content and higher isotopic (13C and 15N) abundance that normally feature mixotrophic orchids because they incorporate N-, 13C-, and 15N-rich fungal biomass. Second, fungal barcoding by next-generation sequencing revealed that the proportion of ectomycorrhizal fungi (expressed as percentage of the total number of either reads or operational taxonomic units) was unusually low compared with E. helleborine growing in situ: instead, we found a high percentage of rhizoctonias, the usual mycorrhizal partners of autotrophic orchids. Altogether, this supports autotrophic survival. Added to the recently published evidence that plastid genomes of mixotrophic orchids have intact photosynthetic genes, this suggests that at least some of them have abilities for autotrophy. This adds to the ecological plasticity of mixotrophic plants, and may allow some reversion to autotrophy in their evolution.